DNA条形码在热带龙脑香科树种鉴定中的应用

胡建霖,刘志芳,慈秀芹,李捷

Use of DNA Barcoding in Identifying Tropical Trees from Dipterocarpaceae

Jianlin Hu,Zhifang Liu,Xiuqin Ci,Jie Li

表2 PCR扩增引物及程序

Table 2 Primer pairs used for PCR amplification and amplification protocol

Barcodes Primers sequence (5'-3') Amplification protocol References rbcL 1F: ATGTCACCACAAACAGAGACTAAAGC 724R: TCGCATGTACCTGCAGTAGC 94°C 4 min; 94°C 30 s, 55°C 45 s, 72°C 1 min, 5 cycles; 94°C 30 s, 54°C 45 s, 72°C 10 min, 30 cycles; 72°C 10 min Fay et al., 1997 Olmstead et al., 1992 matK 3F: CGTACAGTACTTTTGTGTTTACGA 1R: ACCCAGTCCATCTGGAAATCTTGG 94°C 4 min; 94°C 30 s, 51°C 50 s, 72°C 50 s, 35 cycles; 72°C 10 min Unpublished 390F: CGATCTATTCATTCAATATTTC 1326R: TCTAGCACACGAAAGAAGT 94°C 4 min; 94°C 30 s, 50°C 50 s, 72°C 50 s, 35 cycles; 72°C 10 min Cuénoud et al., 2002 trnL-trnF F: GGTTCAAGTCCCTCTATCCC R: ATTTGAACTGGTGACACGAG 94°C 4 min; 94°C 30 s, 55°C 1 min, 72°C 1 min, 35 cycles; 72°C 5 min Taberlet et al.,1991 ITS2 ITS2 2F: ATGCGATACTTGGTGTGAAT ITS2 3R: GACGCTTCTCCAGACTACAAT 94°C 4 min; 94°C 30 s, 55°C 45 s, 72°C 45 s, 35 cycles; 72°C 10 min Chen et al., 2010 Chen et al., 2010 ITS3: GCATCGATGAAGAACGCAGC 26SE: TAGAATTCCCCGGTTCGCTCGCCGTTAC 94°C 4 min; 94°C 30 s, 57°C 45 s, 72°C 45 s, 35 cycles; 72°C 10 min White et al., 1990; Sun et al., 1994